两株四环素降解菌的分离鉴定及降解条件优化

为发掘四环素高效降解菌株,本研究从以四环素为唯一生长碳源的养鸡场粪便堆肥样品中分离筛选到2株四环素降解菌TC-04和TC-09,并通过形态特征、生理生化特征和16SrRNA基因序列分析,对其进行鉴定.采用单因素试验分别探究不同碳源、氮源、微量元素这3个培养基成分和培养时间、接种量和装液量这3个培养条件对两株菌降解四环素...     

饥饿对小规格斑点叉尾鮰体重及鱼体生化组成的影响

将180尾初始体重为（41.9 ± 0.5）g的斑点叉尾鮰放养于18个室内容积为300L的水族箱中进行饥饿处理12周,每箱放10尾,每2周取样一次,研究长期饥饿胁迫下斑点叉尾鮰生长和鱼体生化组成的变化,尤其是肌肉氨基酸和脂肪酸组成的变化。实验结果表明：斑点叉尾鮰的体重、肥满度、脏体比、肝体比和脂体比随着饥饿时间的延长显著降低（p < 0.05）;饥饿过程中全鱼的蛋白质、脂肪含量显著下降（p < 0.05）,水分含量显著增加（p< 0.05）,而灰分含量变化不大;肌肉中蛋白质含量前6周变化不显著（p > 0.05）,而6周后显著降低（p < 0.05）,脂肪、糖原和灰分含量随着饥饿时间的延长显著降低（p < 0.05）,水分含量显著增加（p < 0.05）;肝脏中的蛋白质和水分含量随饥饿时间的延长显著上升（p < 0.05）,粗脂肪和糖原的含量则显著下降（p < 0.05）;在12周的饥饿时间内,总的必需氨基酸所占的百分比变化不显著（p > 0.05）,而总的必需氨基酸和总的非必需氨基酸的比值显著增加（p < 0.05）;斑点叉尾鮰肌肉中总饱和脂肪酸、总单不饱和脂肪酸和n-6 系列脂肪酸的百分含量显著下降（p < 0.05）,而总的n-3系列脂肪酸含量显著增加（p < 0.05）。n-3/n-6脂肪酸的比值随饥饿时间的延长而显著升高（p < 0.05）。以上结果说明：斑点叉尾鮰在饥饿条件下优先动用机体的贮存性物质（脂肪和糖原）作为能源的供给物,而后才被迫动用组织或器官中的功能性物质蛋白质;长期饥饿胁迫下斑点叉尾鮰优先保留必需氨基酸及EPA、DHA等高度不饱和脂肪酸。     

Prognostic Value of LGR5 in Colorectal Cancer: A Meta-Analysis

Objective

Leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5) has recently been reported to be a marker of cancer stem cells (CSCs) in colorectal cancer (CRC), and the prognostic value of LGR5 in CRC has been evaluated in several studies. However, the conclusions remain controversial. In this study, we aimed to evaluate the association between the expression of LGR5 and the outcome of CRC patients by performing a meta-analysis.

Methods

We systematically searched for relevant studies published up to February 2014 using the PubMed, Web of Science, EMBASE and Wangfang databases. Only articles in which LGR5 expression was detected by immunohistochemistry were included. A meta-analysis was performed using STATA 12.0, and pooled hazard ratios (HRs) with 95% confidence intervals (CIs) were used to estimate the strength of the association between LGR5 expression and the prognosis of CRC patients.

Results

A total of 7 studies comprising 1833 CRC patients met the inclusion criteria, including 6 studies comprising 1781 patients for overall survival (OS) and 3 studies comprising 528 patients for disease-free survival (DFS). Our results showed that high LGR5 expression was significantly associated with poor prognosis in terms of OS (HR: 1.87, 95% CI: 1.23–2.84; P = 0.003) and DFS (HR: 2.44, 95% CI: 1.49–3.98; P<0.001). Further subgroup analysis revealed that many factors, including the study region, number of patients, follow-up duration and cutoff value, affected the significance of the association between LGR5 expression and a worse prognosis in patients with CRC. In addition, there was no evidence of publication bias, as suggested by Begg’s and Egger’s tests.

Conclusions

The present meta-analysis indicated that high LGR5 expression was associated with poor prognosis in patients with CRC and that LGR5 is an efficient prognostic factor in CRC.     

Energetics and regulations of formate and hydrogen metabolism by Methanobacterium formicicum

Accumulation of formate to millimolar levels was observed during the growth of Methanobacterium formicicum species on H₂–CO₂. Hydrogen was also produced during formate metabolism by M. formicicum. The amount of formate accumulated in the medium or the amount H₂ released in gas phase was influenced by the bicarbonate concentration. The formate hydrogenlyase system was constitutive but regulated by formate. When methanogenesis was inhibited by addition of 2-bromoethane sulfonate, M. formicicum synthesized formate from H₂ plus HCO _inf3 ^sup- or produced H₂ from formate to a steady-state level at which point the Gibbs free energy (G) available for formate synthesis or H₂ production was approximately -2 to -3 kJ/reaction. Formate conversion to methane was inhibited in the presence of high H₂ pressure. The relative rates of conversion of formate and H₂ were apparently controlled by the G available for formate synthesis, hydrogen production, methane production from formate and methane production from H₂. Results from ¹⁴C-tracer tests indicated that a rapid isotopic exchange between HCOO^- and HCO _inf3 ^sup- occurred during the growth of M. formicicum on H₂–CO₂. Data from metabolism of ¹⁴C-labelled formate to methane suggested that formate was initially split to H₂ and HCO _inf3 ^sup- and then subsequently converted to methane. When molybdate was replaced with tungstate in the growth media, the growth of M. formicicum strain MF on H₂–CO₂ was inhibited although production of methane was not Formate synthesis from H₂ was also inhibited.     

螺环哌啶类似物作为DOR激动剂的特征结构研究

阿片受体激动剂与特定阿片受体亚型结合,常用来治疗与外伤、癌症或心脏病相关的严重疼痛,是十分有吸引力的药用物质．阿片受体有3种经典亚型(δ, κ, μ),均有与其对应的激动剂．δ阿片受体(DOR)激动剂因其还有明显的抗焦虑、抗抑郁和器官保护作用,是非常有前景的药物．本文研究了一批共102个N-取代螺环哌啶类似物作为δ阿片受体激动剂的分子,采用比较分子力场(CoMFA)和比较分子相似性指数(CoMSIA)两种分析方法对所有分子进行了三维定量构效关系(3D-QSAR)研究,其中基于疏水场和氢键供体场参数建立的CoMSIA模型最佳,其模型结果为：Q²=0.501,R²_ncv=0.787,R²_pre=0.780,证明模型自我吻合良好,同时有较强的内部及外部预测能力．而模型的等势线图分析表明,在R₁处引入疏水性的取代基及在R₂处引入亲水性的取代基或氢键供体基团对提高激动剂活性有利．这些结论有助于更好地理解N-取代螺环哌啶类似物作为DOR激动剂的机理,为新型的δ阿片受体激动剂的设计和优化提供一定的指导．     

小麦倒春寒研究现状与进展

由于全球气候变暖,近年来小麦低温灾害事件频发,尤其是拔节-孕穗期的倒春寒灾害已成为制约小麦产量和品质的重要因素之一.本文综述了小麦倒春寒灾害的发生特点(鉴定与分级、时空特征),倒春寒对小麦生理特性(叶片、茎秆、穗部、根系)和产量、质量的影响,总结了抗倒春寒小麦育种、倒春寒危害的分子生物学机制及灾害的监测预警与风险评估等...     

临床常见L型细菌的分布及耐药性变化分析

目的了解临床主要L型致病菌的分布以及对目前常用性变化趋势.方法对2000年7月～2003年7月临床分离的753株药性进行回顾性分析.结果金黄色葡萄球菌、大肠埃希菌、最常见,它们共同对阿米卡星霉素敏感,敏感率达77%以上,共同对罗红霉素、氨苄西林、克林霉素、阿奇霉素耐药,耐药率均在45%以上.结论与我院5年前及10年前L型细菌耐药性对比,抗生素耐药性已发生变迁.掌握L型细菌耐药性的动态,合理使用抗生素.     

Detection of K-ras and p53 mutations in sputum samples of lung cancer patients using laser capture microdissection microscope and mutation analysis

Mutations in the p53 tumor suppressor gene and the K-ras oncogene have been frequently found in sputum and bronchoalveolar lavage (BAL) samples of lung cancer patients and other patients prior to presenting clinical symptoms of lung cancer, suggesting that they may provide useful biomarkers for early lung cancer diagnosis. However, the detection of these gene mutations in sputum and BAL samples has been complicated by the fact that they often occur in only a small fraction of epithelial cells among sputum cells and, in the case of p53 gene, at many codons. In this study, sputum cells were collected on a filter membrane by sputum cytocentrifugation and morphologically analyzed. Epithelial cells were selectively taken by using a laser capture microdissection microscope and analyzed by polymerase chain reaction (PCR) and single-stranded conformational polymorphism (SSCP) for p53 mutations and by PCR and denaturing gradient gel electrophoresis (DGGE) for K-ras mutations. This method was used to analyze sputum of 15 Chinese women with lung cancer from Xuan Wei County, China and detected mutations in sputum of 7 (46.7%) patients, including 5 patients with p53 mutations, 1 patient with a K-ras mutation, and 1 patient with K-ras and p53 mutations. For comparison, only two of the mutations were detected by conventional methods. Therefore, the laser capture/mutation analysis method is sensitive and facilitates the detection of low-fraction mutations occurring throughout the p53 and K-ras genes in sputum of lung cancer patients. This method may be applicable to the analysis of epithelial cells from clinically normal sputum or BAL samples from individuals with a high risk for developing lung cancer.     

Heterozygous Men1 mutant mice develop a range of endocrine tumors mimicking multiple endocrine neoplasia type 1

Multiple endocrine neoplasia type 1 (MEN1) is a hereditary syndrome characterized by the occurrence of multiple endocrine tumors of the parathyroid, pancreas, and anterior pituitary in patients. To study tumorigenesis related to the MEN1 syndrome, we have generated Men1 knockout mice using the gene targeting approach. Heterozygous Men1 mutant mice developed the same range of major endocrine tumors as is seen in MEN1 patients, affecting the parathyroid, pancreatic islets, pituitary and adrenal glands, as well as the thyroid, and exhibiting multistage tumor progression with metastatic potential. In particular, extrapancreatic gastrinoma, pancreatic glucagonoma, and mixed hormone-producing tumors in islets were observed. In addition, there was a high incidence of gonadal tumors of endocrine origin, i.e. Leydig cell tumors, and ovary sex-cord stromal cell tumors in heterozygous Men1 mutant mice. Hormonal disturbance, such as abnormal PTH and insulin levels, was also observed in these mice. These tumors were associated with loss of heterozygosity of the wild-type Men1 allele, suggesting that menin is involved in suppressing the development of these endocrine tumors. All of these features are reminiscent of MEN1 symptoms in humans and establish heterozygous Men1 mutant mice as a suitable model for this disease.     

胃息肉与幽门螺杆菌感染关系研究

目的探讨胃息肉与幽门螺杆菌（Helicobacter pylori,H.pylori）感染关系。方法对1218例胃息肉同时进行H．pylori检查患者进行回顾性分析,分析胃息肉患者H.pylori感染率、胃息肉部位与H.pylori感染关系、胃息肉病理类型与H.pylori感染关系。结果发现胃息肉Hpylori感染患者532例,Hpylori感染率为43．7％。男性胃息肉患者H.pylori感染率为47．5％（216／455）,女性Hpriori感染率感染率为41．4％（316／763）（P〉0．05）,年龄〈20岁、20～39岁、40—59岁和≥60岁胃息肉H.priori感染率分别为41．7％、44．7％、41．6％和47．2％（P〉0．05）;胃窦胃角息肉H.pylori感染率高于其他部位（胃体、胃底和贲门）（P〈0．05）;炎性和增生性胃息肉H.priori感染率高于胃底腺和腺瘤性息肉（P〈0．05）。结论H.pylori感染可能与部分胃息肉发生有一定关系,需要进一步深入研究胃息肉的发生机制。